Genetic element that enhances protein translation
The present invention provides isolated polynucleotides that can serve as translation enhancing elements and their use in protein expression reagents and methods.
1. A recombinant viral or plasmid expression vector comprising
(a) a translation enhancement element (TEE) of between 37 and 42 nucleotides in length, comprising the nucleic acid sequence of 5′-CATATTGAAGAGACAGAGTGATATATAAAACTGCTAA-3′ (SEQ ID NO: 1); and
(b) a cloning site located downstream of the TEE.
2. The expression vector of claim 1 , further comprising a promoter located upstream of the TEE.
3. The expression vector of claim 1 , further comprising a protein encoding nucleic acid cloned into the cloning site.
4. An isolated recombinant host cell comprising the expression vector of claim 1 .
5. A method for protein expression, comprising contacting the expression vector of claim 3 with reagents and under conditions for promoting expression of the protein encoded by the protein-encoding nucleic acid, and expressing the protein.
6. The method of claim 5 , wherein the protein is expressed in a cell-free translation system.
7. The method of claim 5 , wherein the protein is expressed by a recombinant host cell.
8. The method of claim 5 , wherein the expression vector is a recombinant vaccinia virus.
9. The method of claim 7 , wherein the recombinant host cell expresses the protein using non-viral-mediated translation.
10. An isolated recombinant virus, comprising the expression vector of claim 1 , wherein the expression vector is comprises a viral expression vector.
11. The recombinant virus of claim 10 , wherein the recombinant virus is a vaccinia virus, and the viral expression vector is a vaccinia virus expression vector.
12. The expression vector of claim 1 , wherein the TEE consists of 5′-CATATTGAAGAGACAGAGTGATATATAAAACTGCTAA-3′ (SEQ ID NO: 1).
13. The expression vector of claim 1 , wherein the TEE consists of 5′-AGAACCATATTGAAGAGACAGAGTGATATATAAAACTGCTAA-3′ (SEQ ID NO:2).